Skip to Main Content
 

Global Search Box

 
 
 
 

Files

File List

Full text release has been delayed at the author's request until August 18, 2025

ETD Abstract Container

Abstract Header

Interrogation of Cortical Neurogenesis Dysregulation in Autism in iPSC models by Bi-Directional Genome Editing and Lineage Tracing

Fu, Shuai
http://orcid.org/0000-0002-0298-8433
http://rave.ohiolink.edu/etdc/view?acc_num=case1688578849642587

Abstract Details

2023, Doctor of Philosophy, Case Western Reserve University, Molecular Medicine.
Autism Spectrum Disorders (ASDs) are a group of neurodevelopmental disorders for which there is no direct treatment. Understanding the disease mechanisms may lead to future therapeutic targets. In this thesis, we performed bi-directional CRISPR-Cas9 genome editing on induced pluripotent stem cells (iPSCs) derived from individuals with ASD with macrocephaly as well as controls, and uncovered that in addition to PTEN p.Ile135Leu variant, ASD genetic backgrounds also contributed to dysregulating cortical neurogenesis in both 2D neural progenitor cells and 3D cortical organoid models. Surprisingly, ASD specifc PTEN p.Ile135Leu variant dysregulates cortical neurogenesis in an ASD genetic background dependent fashion, as we found that this variant led to overproduction of neural progenitor cell (NPC) subtypes including intermediate progenitor cells (IPCs) and outer radial glia cells (oRGs) as well as neuronal subtypes such as deep and upper layer neurons in the ASD genetic background but not in the control genetic background in the cortical organoids. This study provides strong evidence that both an ASD-specifc PTEN p.Ile135Leu variant and autism genetic background are contributing to the cortical neurogenesis dysregulation. We also developed a lineage tracing system to track neurogenesis in human cortical organoids and applied this system to a control iPSC line as well as autistic isogenic CTNNB1 iPSC lines, we uncovered that the majority of the cortical neurons in the cortical organoids were indirectly generated through IPCs, and neurons derived from diferent lineages were transcriptionally distinct. An ASD-linked CTNNB1 p.Gln76* variant altered the lineage specific production of deep and upper layer neurons as well as the landscape of gene expression profiles among and within different neural progenitor lineages of both deep and upper layer neuron production. Overall, this thesis provides direct evidence that variants in the PTEN and WNT pathways as well as ASD genetic background contribute to dysregulated cortical neurogenesis in ASD-affected individuals.
Anthony Wynshaw-Boris, M.D., Ph.D. (Advisor)
Justin Lathia, Ph.D. (Committee Chair)
Tara DeSilva, Ph.D. (Committee Member)
Fulai Jin, Ph.D. (Committee Member)
Charis Eng, M.D., Ph.D. (Committee Member)
Genetics; Neurosciences
PTEN; ASD genetic background; CRISPR-Cas9 genome editing; iPSC; cortical organoids; Human brain development; neurogenesis; autism spectrum disorders; lineage tracing; single cell RNA-seq; immunohistochemistry.

Recommended Citations

Citations

  • Fu, S. (2023). Interrogation of Cortical Neurogenesis Dysregulation in Autism in iPSC models by Bi-Directional Genome Editing and Lineage Tracing [Doctoral dissertation, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case1688578849642587

    APA Style (7th edition)

  • Fu, Shuai. Interrogation of Cortical Neurogenesis Dysregulation in Autism in iPSC models by Bi-Directional Genome Editing and Lineage Tracing. 2023. Case Western Reserve University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case1688578849642587.

    MLA Style (8th edition)

  • Fu, Shuai. "Interrogation of Cortical Neurogenesis Dysregulation in Autism in iPSC models by Bi-Directional Genome Editing and Lineage Tracing." Doctoral dissertation, Case Western Reserve University, 2023. http://rave.ohiolink.edu/etdc/view?acc_num=case1688578849642587

    Chicago Manual of Style (17th edition)

case1688578849642587
© , all rights reserved.
This open access ETD is published by Case Western Reserve University School of Graduate Studies and OhioLINK.
Release 3.2.12